Quantitative Flow Cytometry to Understand Population Heterogeneity in Response to Changes in Substrate Availability in Escherichia coli and Saccharomyces cerevisiae Chemostats
| Autor: | Heins, Anna-Lena, Johanson, Ted, Han, Shanshan, Lundin, Luisa, Carlquist, Magnus, Gernaey, Krist V., Sørensen, Søren J., Lantz, Anna Eliasson |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Histology
flow cytometry lcsh:Biotechnology Glucose pulse Biomedical Engineering glucose pulse Bioengineering and Biotechnology Bioengineering reporter strain Population heterogeneity membrane robustness Quantitative flow cytometry quantitative flow cytometry lcsh:TP248.13-248.65 Reporter strain Flow cytometry Membrane robustness population heterogeneity Original Research Biotechnology |
| Zdroj: | Frontiers in Bioengineering and Biotechnology, Vol 7 (2019) Heins, A-L, Johanson, T, Han, S, Lundin, L, Carlquist, M, Gemaey, K V, Sørensen, S J & Eliasson Lantz, A 2019, ' Quantitative Flow Cytometry to Understand Population Heterogeneity in Response to Changes in Substrate Availability in Escherichia coli and Saccharomyces cerevisiae Chemostats ', Frontiers in Bioengineering and Biotechnology, vol. 7, 187 . https://doi.org/10.3389/fbioe.2019.00187 Frontiers in Bioengineering and Biotechnology Heins, A-L, Johanson, T, Han, S, Lundin, L, Carlquist, M, Gernaey, K V, Sørensen, S J & Lantz, A E 2019, ' Quantitative Flow Cytometry to Understand Population Heterogeneity in Response to Changes in Substrate Availability in Escherichia coli and Saccharomyces cerevisiae Chemostats ', Frontiers in Bioengineering and Biotechnology, vol. 7, 187 . https://doi.org/10.3389/fbioe.2019.00187 |
| ISSN: | 2296-4185 |
| DOI: | 10.3389/fbioe.2019.00187/full |
| Popis: | Microbial cells in bioprocesses are usually described with averaged parameters. But in fact, single cells within populations vary greatly in characteristics such as stress resistance, especially in response to carbon source gradients. Our aim was to introduce tools to quantify population heterogeneity in bioprocesses using a combination of reporter strains, flow cytometry, and easily comprehensible parameters. We calculated mean, mode, peak width, and coefficient of variance to describe distribution characteristics and temporal shifts in fluorescence intensity. The skewness and the slope of cumulative distribution function plots illustrated differences in distribution shape. These parameters are person-independent and precise. We demonstrated this by quantifying growth-related population heterogeneity of Saccharomyces cerevisiae and Escherichia coli reporter strains in steady-state of aerobic glucose-limited chemostat cultures at different dilution rates and in response to glucose pulses. Generally, slow-growing cells showed stronger responses to glucose excess than fast-growing cells. Cell robustness, measured as membrane integrity after exposure to freeze-thaw treatment, of fast-growing cells was strongly affected in subpopulations of low membrane robustness. Glucose pulses protected subpopulations of fast-growing but not slower-growing yeast cells against membrane damage. Our parameters could successfully describe population heterogeneity, thereby revealing physiological characteristics that might have been overlooked during traditional averaged analysis. |
| Databáze: | OpenAIRE |
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